BIOC 460

DR. TISCHLER
LECTURE 31

 LIPOLYSIS, BETA OXIDATION,

KETONES, LIPOGENESIS
 OBJECTIVES

1. For the lipolytic pathway (lipolysis): describe the pathway, identify

where it occurs, name the principal enzyme involved, and explain the
role of albumin and fatty acid binding protein in the transport and
metabolism of free fatty acids liberated by lipolysis

2. For the degradation of fatty acyl CoAs: describe the roles of acyl CoA
synthetase, carnitine-palmitoyl transferases (CPT-I and CPT-II), and
carnitine acylcarnitine translocase (CAT); discuss the relationship of
products of the -oxidation pathway to energy production.

3. For ketone body metabolism: identify where and when ketone body
formation (ketogenesis) occurs, state the role of ketogenesis, identify
where ketone oxidation occurs and explain why normally individuals
do not develop ketoacidosis even when producing ketone bodies.
 OBJECTIVES

4. Describe the reactions catalyzed by malic enzyme and acetyl CoA

carboxylase

5. For the fatty acid synthase reaction: list the substrates and key products,
identify the sources of NADPH for the reaction, and describe its
general mechanism.

6. Describe how fatty acids are stored as a source of fuel during starvation
or stress.
PHYSIOLOGICAL PREMISE

Would you believe that diabetics having a ketotic crisis

have actually been arrested for DUIs even though they
have consumed no alcohol? Indeed a blood analysis
would show no alcohol. Why would this occur? During a
ketotic crisis a byproduct of the excess ketone production
is acetone. Having nowhere else to go, it is expired
through the lungs. It is the acetone that arresting officers
have smelled on the breath of these individuals and
despite their protestations have innocently believed them
to be consuming alcohol.
 LIPOLYSIS OF STORED TRIACYLGLYCEROL

fatty acids hydrolytically cleaved from triacylglycerol

largely in adipose to release fatty acids as a fuel
may also occur in muscle or liver - smaller amounts of
fatty acids are stored
hormone-sensitive (cyclic AMP-regulated) lipase
initiates lipolysis – cleaves first fatty acid
this lipase and others remove remaining fatty acids
fatty acids/glycerol released from adipose to the
blood
hydrophobic fatty acids bind to albumin, in the blood,
for transport

Triacylglycerol Lipolysis Glycerol + Fatty acids

 CAPILLARY
Lipoproteins
(Chylomicrons L
[2] FABP MITOCHONDRION
or VLDL) P
L FA
acetyl-CoA TCA [7]
A cycle
[3] [4] C
FA -oxidation
FA S [6]
albumin FA FA acyl-CoA acyl-CoA
FABP FABP [5]
FA
carnitine
CYTOPLASM transporter
[1]
fro
m cell membrane
fat FA = fatty acid
cell LPL = lipoprotein lipase
FABP = fatty acid binding protein
ACS = acyl CoA synthetase

Figure 1. Overview of fatty acid degradation

 ATP + CoA AMP + PPi

palmitate palmitoyl-CoA

Cytoplasm

OUTER
ACS MITOCHONDRIAL
CPT-I
[1] [2] MEMBRANE

CoA
palmitoyl-CoA

Intermembrane palmitoyl-carnitine
carnitine
Space
CPT-I defects cause severe muscle weakness because fatty acids are
an important muscle fuel during exercise.

Figure 2 (top). Activation of palmitate to palmitoyl CoA (step 4, Fig.

1) and conversion to palmitoyl carnitine
 CPT-I

palmitoyl-CoA CoA

Intermembrane Space carnitine palmitoyl-carnitine

INNER
CAT [3] MITOCHONDRIAL
MEMBRANE

Matrix CPT-II
carnitine palmitoyl-carnitine
[4]
palmitoyl-CoA CoA

Figure 2 (bottom). Mitochondrial uptake via of palmitoyl-

carnitine via the carnitine-acylcarnitine translocase (CAT)
(step 5 in Fig. 1).
 ATP + CoA AMP + PP
i
Cytoplasm
palmitate palmitoyl-CoA

OUTER
ACS MITOCHONDRIAL
CPT-I
MEMBRANE
[1] [2]

CoA Intermembrane
palmitoyl-CoA Space

carnitine palmitoyl-carnitine

INNER
[3] MITOCHONDRIAL
CAT MEMBRANE

CPT-II
carnitine palmitoyl-carnitine
Matrix
[4]
palmitoyl-CoA CoA
 Palmitoylcarnitine

inner mitochondrial Carnitine

membrane respiratory chain
translocase

Palmitoylcarnitine
matrix side 2 ATP
3 ATP
Palmitoyl-CoA
FAD
oxidation
FADH2 Figure 3.
hydration H2O Processing and
-oxidation of
recycle NAD+ palmitoyl CoA
oxidation
6 times
NADH
thiolase CoA

CH3-(CH)12-C-S-CoA + Acetyl CoA

Citric
O acid
cycle 2 CO2
 Fatty acid 2 Acetyl CoA oxidation to Citric
-oxidation CO2 acid
(excess Thiolase cycle
acetyl CoA) CoA
MITOCHONDRION
Acetoacetyl CoA
acetyl CoA
Figure 4. Ketone body HMG-CoA synthase
formation (ketogenesis)
in liver mitochondria CoA
from excess acetyl CoA Hydroxymethylglutaryl CoA
derived from the -
oxidation of fatty acids HMG-CoA-lyase
acetyl CoA
Acetoacetate
(non-enzymatic) NADH
-Hydroxybutyrate
dehydrogenase
Acetone
NAD+
-Hydroxybutyrate
 KETONE BODY OXIDATION

high rates of lipolysis (e.g., long‑term starvation or in

uncontrolled diabetes) produce sufficient ketones in the
blood to be effective as a fuel
ketones are the preferred fuel if glucose, ketones, fatty acids
all available in the blood
primary tissues: using ketones, when available, are brain,
muscle, kidney and intestine, but NOT the liver.
-Hydroxybutyrate + NAD+  acetoacetate + NADH
-hydroxybutyrate dehydrogenase in mitochondria;
reverse of ketogenesis
 KETOSIS
Excessive build-up of ketone bodies results in ketosis eventually
leading to a fall in blood pH due to the acidic ketone bodies.

Adipose
Tissue X Free fatty Liver
acids

Insulin Ketone Bodies

Pancreas

Figure 5. Mechanism for prevention of ketosis due to

excess ketone body production that can lead to ketoacidosis
LIPOGENESIS
 LIPOGENESIS

 principally in adipose tissue and liver

 lipogenesis – cytoplasm; requires acetyl CoA

 adipose: FA stored as triacylglycerols via esterification

 liver: produces TAG packaged into VLDL and exported

 compounds metabolized to acetyl CoA can serve as a fat

precursor

 glucose = primary source of carbons for fat synthesis.

 Glucose CYTOPLASM MITOCHONDRIAL MATRIX

PPP Glycolysis NAD, CoA NADH, CO2

Fatty
Acids PDH
CO2 Pyruvate Pyruvate Acetyl
NADPH ATP, CO2 CoA
ME PC
FAS
Malate NADP +

Malonyl CoA NAD+ ADP, Pi

ADP, Pi MDH Oxaloacetate CS
ACC NADH
CO2, ATP Oxaloacetate
Acetyl CoA ADP+Pi

ATP, CoA
CL
Citrate Citrate

Figure 6. Export of acetyl CoA as citrate for fatty acid biosynthesis,

generation of NADPH and pathway of lipogenesis. (similar to diagram
discussed for cholesterol synthesis exxept this involves PDH reaction)
 KEY MITOCHONDRIAL REACTIONS

PYRUVATE CARBOXYLASE

pyruvate + CO2 + ATP  oxaloacetate + ADP + Pi

PYRUVATE DEHYDROGENASE

pyruvate + NAD+ + coenzyme A (CoA)  acetyl CoA + CO2 + NADH

 KEY CYTOPLASMIC REACTIONS
INDIRECTLY NEEDED FOR LIPOGENESIS

Citrate Lyase
citrate + CoA + ATP  acetyl CoA + oxaloacetate + ADP + Pi

Malate dehydrogenase
oxaloacetate + NADH  malate + NAD+

Malic Enzyme
malate + NADP+  pyruvate + NADPH
 KEY CYTOPLASMIC REACTIONS DIRECTLY NEEDED
FOR LIPOGENESIS AND FATTY ACID ACTIVATION

Acetyl CoA Carboxylase:

acetyl CoA + HCO3- + ATP  malonyl CoA + ADP + Pi

Fatty Acid Synthase:

acetyl CoA + 7 malonyl CoA + 14 NADPH + 14 H+ 
palmitate + 7 CO2 + 8 CoA + 14 NADP+

Acyl CoA Synthetase:

(also used for fatty acids other than palmitate)
palmitate + ATP + CoA  palmitoyl CoA + AMP + PPi
 reduction
dehydration A
A A reduction
condensation CE C
CE C CE C acp P
acp P acp P
CO2 2 NADPH 2 NADP+ C=O
C=O C=O CO2 C=O
CH2
CHC=O CH2 CO2
C=O CH2
3
C=O C=O CH2
CH3C=O - CO2
COO CH CH2 CH2 C=O
CH 2
CH3
acetyl malonyl
3
C=O C=O C=O CH3
CoA CoA
CH3 CH3 CH3 4-C
unit
Figure 7. General mechanism for the fatty acid synthase
reaction. CE is condensing enzyme. ACP is acyl carrier
protein. This row represents the initial steps for priming the
reaction with acetyl CoA and the addition of two carbons from
malonyl CoA.
 reduction
dehydration A
A A reduction
condensation CE C
CE C CE C acp P
acp P acp P
CO2 2 NADPH 2 NADP+
6-C
4-C malonyl 6-C
unit
unit CoA unit

Figure 7. General mechanism for the fatty acid synthase

reaction. CE is condensing enzyme. ACP is acyl carrier
protein. This row depicts a typical cycle of adding two more
carbons to the fatty acid chain.
 A 5 more cycles A
CE C adding 10 CE C
acp P more carbons acp P
thioesterase
cleavage
6-C malonyl 5malonyl CoA
5CO2 16-C
unit CoA 10NADPH unit
10NADP+
palmitate
A
CE C
palmitate acp P

Figure 7. General mechanism for the fatty acid synthase

reaction. CE is condensing enzyme. ACP is acyl carrier
protein. This row shows the release of the finished product,
palmitate, through cleavage by thioesterase.
Sources of NADPH for the Biosynthesis of Fatty Acids.

malic enzyme:

Malate + NADP+  Pyruvate + CO2 + NADPH

pentose phosphate pathway:

Glucose-6-P + 2 NADP+  Ribulose-5-P + 2

NADPH + CO2
 Glycerol
Dihydroxyacetone phosphate
ATP glycerol fatty acyl CoA
kinase
ADP
Glycerol-3-P CoA
fatty acyl CoA Acyldihydroxyacetone phosphate
NADPH
CoA
Lysophosphatidic NADP+
acid fatty acyl CoA
Pi
CoA
Phosphatidic Diacylglycerol
acid phosphatase fatty acyl CoA

CoA
Triacylglycerol
Figure 8. Formation of phosphatidic acid from glycerol-3-P or
DHAP, and its conversion to triacylglycerol